Download Protocol S3: 96-well yeast liquid growth assays

Protocol S1: 96-well yeast liquid growth assays
Individual yeast transformants carrying the designated plasmids were inoculated
into single wells of a 96-well plate containing non-inducing selective synthetic media
supplemented with 2% glucose. Plates were incubated 12-18 hours. Yeast were then
transferred to a solid media tray containing non-inducing selective synthetic media
supplemented with 4% glucose with a 3.18 mm diameter 96 floating pin tool (V&P
Scientific, Inc., San Diego, CA). After one to two days, the yeast spots on the solid plates
were transferred to fresh non-inducing liquid media containing 96-well plates using a
1.58 mm diameter 96 floating pin tool and incubated 16-18 hours such that yeast grew to
OD600 between 0.3-0.5. The liquid cultures were then transferred with a 1.58 mm
diameter floating pin tool into liquid 96-well plates containing inducing selective
synthetic media (2% galactose) or inducing rich media ((YEP plus clonNAT (70 (g/ml)
(Werner BioAgents, Jena, Germany) plus 2% galactose)). Plates were incubated for 48
hours without agitation and growth was measured by OD600 readings using a Victor II
Plate Reader (Perkin Elmer, Wellesley, MA). All incubations were done at 30°C.